Manufactured seed having parabolic cavity

ABSTRACT

The present disclosure includes a manufactured seed comprising a seed shell and a restraint disposed within the seed shell. The seed shell is a structure having an open end and a closed end. The restraint comprises a parabolic cavity. A seal assembly is disposed on the open end of the seed shell in a configuration effective to seal the seed shell.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is entitled to and claims the benefit of priority under35 U.S.C. §119 from U.S. Provisional Patent Application Ser. No.61/502,726 filed Jun. 29, 2011, and titled “Manufactured Seed HavingParabolic Cavity,” the contents of which are incorporated herein byreference.

This application relates to the following applications, filed on thesame day as the present patent application, the contents of which areall incorporated herein by reference:

U.S. patent application Ser. No. 13/533.260 and titled “Shoot Restraintfor Use With Manufactured Seeds;”

U.S. patent application Ser. No. 13/533,486 and titled “ManufacturedSeed Having Parabolic End Seal Assembly;” and

U.S. patent application Ser. No. 13/533,287 and titled “ManufacturedSeed Having Parabolic End Seal Assembly and Parabolic Cavity;”

U.S. patent application Ser. No. 13/533,213 and titled “ManufacturedSeed Having Treated End Seal Assembly;” and

U.S. patent application Ser. No. 13/533,540 and titled “ManufacturedSeed Having Embryo Disposed Therein.”

TECHNICAL FIELD

The present disclosure is directed generally to improved designs formanufactured seeds and manufactured seed components.

BACKGROUND

Modern research leading to the successful demonstration of encapsulationof tissue culture derived from plants has initiated a line of researchfocused on the development of synthetic or “manufactured” seeds.Manufactured seeds generally include encapsulated somatic or zygoticplant embryos that functionally mimic development of naturallypropagated seeds. Such manufactured seeds may reduce labor costs andincrease efficiency in many modern agriculture (including silviculture)applications. Examples of manufactured seeds are disclosed, for example,in U.S. Pat. No. 5,701,699, issued to Carlson et al., the disclosure ofwhich is hereby expressly incorporated by reference.

Typical manufactured seeds include a seed shell, synthetic gametophyte,and a plant embryo. A manufactured seed that does not include the plantembryo is known in the art as a “seed blank.” The seed blank typicallyis a cylindrical capsule having a closed end and an open end. Thesynthetic gametophyte is placed within the seed shell to substantiallyfill the interior of the seed shell. A longitudinally extending hardporous insert, known as a shoot restraint, may be centrally locatedwithin one end of the seed shell, surrounded by the syntheticgametophyte, and includes a centrally located cavity extending partiallythrough the length of the shoot restraint. The plant embryo is depositedwithin the cavity of the shoot restraint. The plant embryo is thensealed within the seed blank by an end seal, which may be coated with anantibiotic substance.

Although known manufactured seeds are generally effective in providingan inexpensive delivery unit for plant tissue culture, there are manyopportunities to improve current seed design. For example, one problemwith some manufactured seeds involves low numbers of successfulgerminants. Many factors can lead to germination failure; however, mostmanufactured seeds typically exhibit some form of abnormal growth thatindicates germination may not be successful. Being able to identifyabnormal growth patterns and provide seed design solutions couldsignificantly help advance manufactured seed technology.

Thus, there is a need to continually improve the design of manufacturedseeds to reduce abnormal growth of embryos. Improvements to manufacturedseed design that lead to an increased number of successful and vigorousgerminants (when compared with conventional manufactured seed designs)would be particularly desirable in this field of developing technology.Ideally, seed design improvements may also help improve cost andefficiency models for utilization of manufactured seed technology.

SUMMARY

The following summary is provided for the benefit of the reader only andis not intended to limit in any way the invention as set forth by theclaims. The present disclosure is directed generally towards improveddesigns for manufactured seeds and manufactured seed components.

The present disclosure includes a manufactured seed comprising a seedshell and a restraint disposed within the seed shell. The seed shell isa structure having an open end and a closed end. The restraint comprisesa parabolic cavity. A seal assembly is disposed on the open end of theseed shell in a configuration effective to seal the seed shell.

BRIEF DESCRIPTION OF THE DRAWINGS

The present disclosure is better understood by reading the followingdescription of non-limitative embodiments with reference to the attacheddrawings wherein like parts of each of the figures are identified by thesame reference characters, and are briefly described as follows:

FIG. 1 is a side cross-sectional view of an embodiment of a manufacturedseed according to the disclosure;

FIG. 2 is a side cross-sectional view of another embodiment of amanufactured seed according to the disclosure;

FIG. 3 is a perspective view of a seed shell according to embodiments ofthe disclosure;

FIG. 4 is a top plan view of the seed shell from FIG. 3;

FIG. 5 is a side cross-sectional view of the seed shell from FIGS. 3 and4;

FIG. 6 is a side cross-sectional view of another embodiment of amanufactured seed according to the disclosure;

FIG. 7 is a side cross-sectional view of another embodiment of amanufactured seed according to the disclosure;

FIGS. 8 and 9 are perspective views of an embodiment of a restraintaccording to the disclosure;

FIG. 10 is a side cross-sectional view of the restraint from FIGS. 8 and9;

FIG. 11 is a top plan view of the restraint from FIGS. 8-10;

FIGS. 12 and 13 are perspective views of another embodiment of arestraint according to the disclosure;

FIG. 14 is a side cross-sectional view of the restraint from FIGS. 12and 13;

FIG. 15 is a top plan view of the restraint from FIGS. 12-14;

FIGS. 16-21 are side cross-sectional views of embodiments of amanufactured seed according to the disclosure;

FIGS. 22 and 23 are perspective views of embodiments of end sealassemblies according to embodiments of the disclosure;

FIGS. 24-26 are side views of cavities having embryos disposed thereinaccording to embodiments of the disclosure;

FIG. 27 is a bar chart showing breaking strength of seal assembliestreated with conventional coatings;

FIG. 28 is a bar chart showing breaking strength of seal assembliestreated according to embodiments of the disclosure and conventional sealassemblies;

FIG. 29 is a bar chart showing percent population of partial germinantsand full germinants in an Example;

FIGS. 30 and 31 are bar charts showing breaking strength of sealassemblies treated according to embodiments of the disclosure;

FIG. 32 is a bar chart showing breaking strength of seal assembliesaccording to embodiments of the disclosure and conventional sealassemblies;

FIG. 33 is a bar chart showing effectiveness of cotyledon extraction formanufactured seeds according to embodiments of the disclosure andconventional seeds;

FIG. 34 is a bar chart showing germinants with extraction issues formanufactured seeds according to embodiments of the disclosure andconventional seeds; and

FIG. 35 is a bar chart showing normalcy for manufactured seeds accordingto embodiments of the disclosure and conventional seeds.

DETAILED DESCRIPTION

The present disclosure describes improved designs for manufactured seedsand manufactured seed components. Certain specific details are set forthin the following description and FIGS. 1-35 to provide a thoroughunderstanding of various embodiments of the disclosure. Well-knownstructures, systems, and methods often associated with such systems havenot been shown or described in detail to avoid unnecessarily obscuringthe description of various embodiments of the disclosure. In addition,those of ordinary skill in the relevant art will understand thatadditional embodiments of the disclosure may be practiced withoutseveral of the details described below. Certain terminology used in thedisclosure are defined as follows:

“Agricultural crop” refers to cultivated plants or agricultural producesuch as grain, vegetables, or fruit. As non-limiting examples,agricultural crops according to the disclosure include corn, soybean,rice, wheat, sugar cane, canola, coffee, banana, and cotton.

“Cotyledon” refers generally to the first, first pair, or first whorl(depending on the plant type) of leaf-like structures on the plantembryo that function primarily to make food compounds in the seedavailable to the developing embryo but in some cases act as food storageor photosynthetic structures.

“Dead end” refers to the closed end of a manufactured seed.

“Dimple” refers to depressions, indentations, or protrusions in asurface of a material. Examples of various types of dimples according toembodiments of the disclosure are illustrated in the Figures.

“Epicotyl” refers to the portion of the plant developed aftergermination from the stem apex.

“Functional contact” with respect to orientation of embryos according tothe disclosure is intended to mean in a position in which the embryo isconfigured to uptake nutrients from the nutritive media.

“Germinant” means an embryo that has undergone sufficient growth anddevelopments to protrude from the seed shell of a manufactured seed.This stage is generally analogous to protruding from a natural botanicseed.

“Hypertrophy” refers to a type of abnormal growth characterized byabnormally large or swollen portions of the embryo.

“Hypocotyl” refers to the portion of the plant embryo or seedlinglocated below the cotyledons but above the radicle.

“Live end” refers to the open end of a manufactured seed.

“Nutritive media” refers to a source of nutrients, such as vitamins,minerals, carbon, and energy sources, and other beneficial compoundsused by the embryo during germination.

“Parabolic” refers to a geometric configuration having at least oneparabolic cross-sectional area and at least one elliptical and/orcircular cross-sectional area. Examples of parabolic configurationsaccording to embodiments of the disclosure are illustrated in theFigures.

“Paraffin oil” refers to mineral oil or any other type of oil havingheavier alkanes (a density of approximately around 0.8 g/cm³) including,but not limited to nujol, adepsine oil, alboline, glymol, medicinalparaffin, or saxol.

“Radicle” refers to the part of a plant embryo that develops into theprimary root of the resulting plant.

“Root end” or “root portion” with respect to a plant embryo refers tothe portion of the embryo from which the non-aerial part of the plantoriginates.

“Shoot end” or “shoot portion” with respect to a plant embryo refers tothe portion of the embryo from which the aerial part of the plantoriginates.

“Somatic embryo” is a plant embryo that developed via laboratoryculturing of totipotent plant cells or by induced cleavage polyembryony.

“Tree” refers to any type of woody perennial plant. The disclosure isnot intended to be limited to a particular species or type of tree.

“Zygotic embryo” refers to a plant embryo originating from a naturalseed of the corresponding plant.

Overview

Referring to FIGS. 1, 2, 6, 7, 16-21 a manufactured seed 10 constructedin accordance with embodiments of the disclosure is shown arranged on acoordinate system comprising an x-axis (X), a y-axis (Y), and a z-axis(Z). FIGS. 3-5, 8-15, 22-26 depict details of embodiments of variouscomponents of the manufactured seeds 10. FIGS. 1, 6, 16, 18, and 20generally illustrate embodiments of manufactured seeds that areparticularly useful in applications involving tree embryos. FIGS. 2, 7,17, 19, and 21 generally illustrate embodiments of manufactured seedsthat are particularly useful in applications involving embryos fromagricultural crops. However, a person of ordinary skill in the art willappreciate that manufactured seeds having a structure or design that issubstantially similar to those shown in FIGS. 1, 6, 16, 18, and 20 maybe used with agricultural crop embryos. Similarly, manufactured seedshaving a structure or design that is substantially similar to thoseshown in FIGS. 2, 7, 17, 19, and 21 may be used with tree embryos.

Referring to FIGS. 1 and 2, manufactured seeds 10 according toembodiments of the disclosure generally include the followingcomponents: a seed shell 12, a shoot restraint 14 disposed within theseed shell 12, and a seal assembly 16 (e.g., a live end seal). The shootrestraint 14 includes a longitudinally extending cavity 18 that extendsat least partially through the length of the shoot restraint 14. Anembryo 20 may be disposed in the cavity 18.

Manufactured seeds according to embodiments of the disclosure includeseed design features intended to promote healthy and vigorousgermination. Seed design features according to embodiments of thedisclosure are based on identifying and inhibiting abnormal growth. Forexample, some studies have shown that one type of abnormalgrowth—hypertrophy—can limit axial elongation of the embryo, therebyresulting in failure to shed the seed shell. Thus, manufactured seedsaccording to embodiments of the disclosure have been modified to reducethe mechanical resistance (e.g., including but not limited tohypertrophy) that can cause abnormal growth patterns or inhibitgermination for embryos that may already exhibit abnormal growthpatterns.

There are many other examples of abnormal growth patterns that mayindicate a higher potential for germination failure. Some studies showthat during development, embryos may become stuck on the seal assemblyand/or open end of the seed shell while exiting the manufactured seed.In other situations, germination may be inhibited because the breakingstrength of the seal assemblies are too weak or too strong. If thebreaking strength is too strong, the seal assemblies do not break andallow the growing embryo to emerge from the seed shell. On the otherhand, if the end seals are too weak, seal assemblies can ruptureprematurely before the embryo is sufficiently developed, therebyinhibiting germination. In addition, some evidence suggests thatconventional coatings for end seals (e.g. triple antibiotic ointment)are possibly detrimental to germination. Features of manufactured seeds10 according to embodiments of the disclosure, including features oftheir various components, are described in further detail below.

Seed Shell

FIGS. 3-5 illustrate details of seed shells 12 according to embodimentsof the disclosure. In some embodiments, the seed shell 12 may besuitably formed from a section of tubular material. In otherembodiments, the seed shell may spherical, ovaloid, cubical, or anyother shape that would be suitable to a person of ordinary skill in theart. As shown In FIGS. 1-3 and 5, the seed shell 12 has an open end 22(e.g., a live end) and a closed end 24 (e.g., a dead end seal). As shownin FIGS. 1 and 2, the closed end 24 may be sealed using an end seal 28or any other means known to a person of ordinary skill in the art.

Seed shells 12 according to embodiments of the disclosure may befabricated from a variety of materials including, but not limited to,cellulosic materials, glass, plastic, moldable plastic, cured polymericresins, paraffin, waxes, varnishes, and combinations thereof such as awax-impregnated paper. The materials from which the seed shell 12 ismade are generally non-toxic and provide a degree of rigidity. The seedshell 12 can be biodegradable, although typically the seed shell remainsintact and resistant to degradation until after emergence of thegerminating embryo.

Referring to FIGS. 1 and 2, the seed shell 12 may be configured to housea nutritive medium 26 that is in functional contact with the embryo 20.Nutritive media 26 according to the disclosure may include a substancethat causes the media to be a semi-solid or have a congealed consistencyunder normal environmental conditions. Suitable nutritive media 26 aredescribed, for example, in U.S. Pat. No. 5,701,699 and U.S. PatentApplication Ser. No. 61/387,244, the disclosures of which are herebyincorporated by reference.

As shown in FIGS. 3-5, seed shells 12 according to embodiments of thedisclosure may have numerous different dimensions for use with differenttypes of embryos. In some embodiments, the seed shell 12 has a seedshell diameter D1 ranging about 0.20 inches to about 0.30 inches. Inother embodiments the seed shell diameter D1 may be about 0.30 inches toabout 0.40 inches. The seed shell 12 also has a seed shell length L1 anda seed shell thickness T1. L1 may be approximately 0.75 inches toapproximately 1.25 inches. T1 may be approximately 0.015 inches toapproximately 0.030 inches.

Shoot Restraint

As described above, manufactured seeds 10 according to embodiments ofthe disclosure include a shoot restraint 14 disposed longitudinallywithin the seed shell 12. In some embodiments, the shoot portion of theembryo includes cotyledons. Sometimes the cotyledons may be removed fromthe shoot portion of the embryo prior to being disposed in the shootrestraint 14. In some cases, the shoot portion of the embryo does nothave cotyledons. The shoot restraint 14 generally functions todiscourage abnormal growth by preventing the shoot portion of the embryofrom becoming trapped. FIGS. 6-15 illustrate details of shoot restraints14 according to embodiments of the disclosure.

FIGS. 6 and 7 are side cross-sectional views of manufactured seedsaccording to embodiments of the disclosure without seal assemblies orembryos shown to better illustrate embodiments of suitable shootrestraints. In some embodiments, the shoot restraint 14 is centered toextend longitudinally into the seed shell 12. Shoot restraints 14 may bemanufactured from a porous material having a hardness strong enough toresist puncture or fracture by a germinating embryo. Suitable materialsinclude ceramic, porcelain, or other similar materials known to a personof ordinary skill in the art. The shoot restraint 14 generally includesa longitudinally extending cavity 18 that is configured to receive anembryo.

Shoot restraints 14 according to embodiments of the disclosure areconfigured to promote normal germination of embryos by providing a shapethat facilitates healthy emergence of the embryos from the manufacturedseed. Further, cavities 18 in the shoot restraints 14 according toembodiments of the disclosure have been designed to facilitate alternateorientations of embryos and to promote healthy germination analogous tothat of a natural botanical seed. Specifically, cavities according toembodiments have a substantially parabolic shape that is expected tohelp promote germination by reducing the likelihood of abnormal growth.

FIGS. 8-15 illustrate further details of embodiments of shoot restraints14 and cavities 18 according to the disclosure. In some embodiments,shoot restraints 14 include a substantially cylindrical upper portion30, a substantially cylindrical lower portion 32, a top surface 34, abottom surface 36, and a restraint depth L2. The top surface 34 and thebottom surface 36 have a substantially circular cross-section. A cavity18 having a substantially parabolic shape extends from the top surface34 to the bottom surface 36 through the upper portion 30 and at leastpartially into the lower portion 32. In some embodiments, the bottomsurface 36 is chamfered. In other embodiments, the bottom surface 36 maybe flat or rounded.

FIGS. 8-11 depict embodiments of shoot restraints 14 according to thedisclosure similar to the restraint shown in FIG. 6. FIGS. 12-15 depictembodiments of shoot restraints 14 according to embodiments of thedisclosure similar to the restraint shown in FIG. 7. Generally, FIGS.8-11 depict a narrower and deeper shoot restraint 14 when compared tothe restraints shown in FIGS. 12-15. Table 1 below summarizes suitableranges for dimensions of the various components in restraints accordingto embodiments of the disclosure. Further, a person of ordinary skill inthe art will appreciate that the restraints shown may be modified toinclude slightly different dimensions and configurations withoutdeparting from the spirit of the disclosure.

TABLE 1 Exemplary Dimensions for Restraints in FIGS. 8-15 Value in Rangefor Value in Range for FIGS. Dimension FIGS. 8-11 FIGS. 8-11 FIGS. 12-1512-15 L2 0.261 inches  0.20 to 0.30 inches 0.200 inches  0.15 to 0.30inches L3 0.063 inches  0.20 to 0.30 inches 0.063 inches  0.15 to 0.30inches L4 0.198 inches 0.10 to .0250 inches 0.137 inches  0.10 to 0.15inches L5 0.035 inches  0.01 to 0.05 inches 0.035 inches  0.01 to .05inches L6 0.028 inches  .001 to .05 inches 0.028 inches  0.01 to 0.05inches L7 0.118 inches 0.09 to 0.220 inches 0.118 inches 0.09 to 0.220inches D2 0.240 inches  0.15 to 0.30 inches 0.325 inches  0.20 to 0.35inches D3 0.214 inches  0.15 to 0.29 inches 0.299 inches  0.25 to 0.35inches D4 0.147 inches  0.10 to 0.18 inches 0.140 inches  0.10 to 0.16inches D5 0.144 inches  0.10 to 0.20 inches 0.234 inches  0.10 to 0.30inches θ    2 degrees      0 to 5 degrees    2 degrees      0 to 5degrees Ψ    5 degrees      0 to 7 degrees    5 degrees      0 to 7degrees Φ   20 degrees     0 to 25 degrees   20 degrees     0 to 25degrees

Referring to FIGS. 8-15, the upper portion 30 has an upper depth L3 andthe lower portion 32 has a lower depth L4, the lower depth L4 beinglarger than the upper depth L3. In some embodiments, the upper portion30 of the shoot restraint 14 may have a single diameter. Referring toFIGS. 11 and 15, the upper portion 30 may comprise an upper portiondiameter D2. In other embodiments, the upper portion 30 of the shootrestraint 14 comprises one or more sections having varying diameters.For example, referring to FIGS. 10 and 14, the upper portion 30 maycomprise a first section 38 (having a first section depth L5) and asecond section 40 (having a second section depth L6). In someembodiments, the sidewalls of the first section 38 and the secondsection 40 are chamfered or angled with respect to the Y axis asdepicted in the Figures. In other embodiments, the sidewalls of theupper portion 30 are substantially straight. In some embodiments, theupper portion 30 comprises an edge 42 having an edge diameter D3.

The lower portion 32 of the shoot restraint 14 may have sidewalls thatare substantially straight with respect to the Y-axis or sidewalls thatare substantially angled as shown in the Figures. In some embodiments,the sidewalls are tapered such that the lower portion 32 has a lowerportion diameter D4 than is substantially smaller than the upper portiondiameter D2.

Referring to FIGS. 8-15, the cavity 18 extending from the top surface 34to the bottom surface 36 has a substantially parabolic shape and acavity diameter D5 that varies along the length of the cavity 18. At itswidest point, D5 may be about the same size as the upper portiondiameter D2. In other embodiments, D5 may be slightly larger or smallerthan the specific values disclosed provided that D5 does not exceed D2.Cavities 18 according to embodiments of the disclosure extend past theupper portion 30 and at least partially into the lower portion 32 asshown by a cavity depth L7.

Seal Assembly

As described above, manufactured seeds 10 according to embodiments ofthe disclosure include a seal assembly 16 disposed on the seed shell 12to effectively seal the seed shell. FIGS. 16-23 depict variousembodiments of seal assemblies 16 according to the disclosure. Referringto FIGS. 16-21, the seal assembly 16 includes a primary end seal 44arranged on the open end 22 of the seed shell 12. The primary end seal44 may suitably formed from biodegradable plastic and includes acentrally located opening 52. The opening 52 is sized to correspond tothe diameter of the cavity 18 (D5) to permit a germinating embryo (notshown in this Figure) to pass through. The primary end seal 44 may besuitably attached by a variety of well-known methods, including glue orheat bonding.

A secondary end seal 46 may be arranged on the primary end seal 44. Thesecondary end seal 46 may be suitably formed from a well-known selfsealing, moldable, and flexible film (commercially available asParafilm®). The secondary end seal 46 may be formed and attached to theprimary end seal 44 by a well-known method, such as heat bonding orgluing. In some embodiments, a sealing wax may be used to facilitatebonding between the primary end seal 44 material and the film of thesecondary end seal 46. In some embodiments, the secondary end seal 46includes one or more dimples 50, each having a substantially parabolicshape. The properties of dimples 50 according to embodiments of thedisclosure will be described in further detail below.

Referring to FIGS. 22 and 23, secondary end seals 46 according toembodiments of the disclosure are shown having outwardly extendingdimples 50 a and inwardly extending dimples 50 b. Embodiments ofsecondary end seals 46 according to the disclosure may comprise a singledimple or more than one dimple. For example, as shown in FIG. 22, thesecondary end seal 26 includes a single outwardly extending dimple 50 a.In other embodiments, the secondary end seal 46 may comprise more thanone dimple (e.g. FIG. 23). Some of the dimples may extend inwardly whileothers may extend outwardly.

Dimples 50 according to the disclosure may be formed by using a suitablyshaped steel pin (or another equivalent device) to stretch theParafilm®. Pins used according to methods in the disclosure may beparabolic, ellipsoid, hemispherical, or any other suitable shape knownto a person of ordinary skill in the art. The stretching results in asubtle loss of breaking strength in the stretched areas of the secondaryend seal 46. In some embodiments, the resulting loss in breakingstrength may be anywhere from about 10% to about 30%. Accordingly, whenthe embryo emerges from the seed shell 12, it is exposed to a secondaryend seal 46 having a non-uniform breaking strength across its surface.

Referring to FIGS. 22 and 23, the dimple or dimples 50 on the secondaryend seal 46 define a pre-stressed area 54 that has a lower breakingstrength than the rest of the dimple 50 (e.g., the non-pre-stressed area55). In some embodiments, the pre-stressed area 54 has a pre-stresseddiameter D7 that is at least as wide as the cavity diameter D5. In someembodiments, D7 may be about 50% to about 95% as wide as D5. In somecases, seal assemblies 16 having a pre-stressed area 54 that is almostas wide or significantly wider than the cavity diameter D5 may beparticularly helpful in promoting germination. For example, D7 may beabout 80% to about 300% as wide as D5.

Referring back to FIGS. 16-21, secondary end seals 46 according toembodiments of the disclosure may include dimples 50 having variousdimple depths L8 and dimple diameters D6. In some embodiments (e.g.,FIG. 17) dimple diameters D8 can be about the same size as the seedshell diameter D1. In other embodiments, dimple diameters D8 aresmaller. Dimple depths L8 may be as deep as 0.04 inches. Outwardlyextending dimple depths L8 are measured according to the distance thatthe dimple 50 extends away from the closed end 24 of the seed shell 12.Inwardly extending dimple depths L8 are measured according to thedistance that the dimple 50 extends towards the closed end 24 of theseed shell 12. Referring specifically to FIG. 21, in some embodiments,dimples 50 according to embodiments of the disclosure may extend pastthe primary end seal 44 towards the closed end 24 of the seed shell. Insome cases, a first portion of the secondary end seal 46 is disposed onone side of the primary end seal 44 while a second portion of thesecondary end seal 46 remains on the opposite side of the primary endseal 44.

A person of ordinary skill in the art will appreciate that a singlesecondary end seal 46 may have a single dimple 50 or more than onedimple 50. If the secondary end seal 46 has more than one dimple 50, thedimensions of each dimple 50 may be uniform or may vary substantially.Further, a single secondary end seal 46 may comprise both inwardlyextending dimples 50 a and outwardly extending dimples 50 b.

In some embodiments, a tertiary end seal 48 may be arranged on thesecondary end seal 46. The tertiary end seal 48 may be made from amaterial that degrades in structural integrity after a predeterminedexposure to environmental conditions. The tertiary seal 48 also servesas an anti-microbial barrier to protect around the live end of themanufactured seed as the embryo germinates and emerges from within theseed shell 12. Suitable materials used to manufacture the tertiary seals48 include water soluble materials, wax, environmentally degradablematerials, and biodegradable materials.

Treated Seal Assemblies

Seal assemblies 16 may be treated with a substance to improvegermination of embryos 20 within the seed shell 12. Paraffin oil hasbeen found to be a suitable treatment due to its ability to softenportions of the sealing assembly 16. In some embodiments, the secondaryend seal 46 is coated with a paraffin oil in a quantity sufficient toreduce the breaking strength of the secondary end seal 46. This amountmay vary depending on whether the secondary end seal 46 has beenpre-stressed according to some embodiments described above. In someembodiments, the secondary end seal is not pre-stressed and paraffin oilis applied. The surface of the secondary end seal 46 may be partiallycoated with paraffin oil or the entire surface of the secondary end seal46 may be covered with the paraffin oil.

Varying amounts of paraffin oil may be used according to embodiments ofthe disclosure. Too little paraffin oil may be insufficient to reducebreaking strength and encourage germination. However, too much paraffinoil may cause root damage. In some embodiments, a suitable range ofparaffin oil is approximately 0.5 mg to approximately 10.0 mg. In otherembodiments, a suitable range of paraffin oil is approximately 3.0 mg toapproximately 6.0 mg.

Different types of paraffin oils may be suitable for use withembodiments according to the disclosure. In a non-limiting example,suitable paraffin oils have a density of approximately 0.827 g/mL toapproximately 0.890 g/mL at 20° C. In another non-limiting example,suitable paraffin oils have a dynamic viscosity approximately 110 mPasto approximately 230 mPas. Paraffin oils according to the disclosure maybe in the form of viscous liquids or may include solids.

Embryo Orientation

As described above, an embryo 20 may be disposed in the cavity 18 ofmanufactured seeds according to embodiments of the disclosure. FIGS.24-26 illustrate different orientations of embryos in shoot restraints14 having cavities 18 according to embodiments of the disclosure. Eachembryo 20 includes a shoot end 56 and a root end 58. In someembodiments, the shoot end 56 may comprise one or more cotyledons if thetype of embryo used in the manufactured seed has cotyledons. In someembodiments, cotyledons may be removed from the shoot end 56 of theembryo 20 prior to inserting the embryo 20 into the cavity 18.

Generally the embryo 20 is oriented shoot end 56 first. FIG. 24 shows anembodiment in which the embryo 20 is disposed in a perpendicularorientation with respect to a plane 60 in which the sealing assembly(not shown in the Figure) is arranged. FIG. 25 shows an embodiment inwhich the embryo 20 is disposed in a parallel orientation with respectto the plane 60. FIG. 26 shows an embodiment in which the embryo 20 isdisposed in a skew or sloped orientation with respect to a plane 60. InFIG. 26, a slope angle β is shown defined by the embryo 20 and theY-axis or a sidewall 62 (see, e.g., FIGS. 1, 2, 6, and 7) of the seedshell 12. The slope angle β may be less than 90 degrees. In somepreferred embodiments, the slope angle β is about 45 degrees to about 70degrees.

Words in the above disclosure using the singular or plural number mayalso include the plural or singular number, respectively. For example,the term “dimple” could also apply to “dimples.” Additionally, the words“herein,” “above,” “below” and words of similar import, when used inthis application, shall refer to this application as a whole and not toany particular portions of this application.

From the foregoing, it will be appreciated that the specific embodimentsof the disclosure have been described herein for purposes ofillustration, but that various modifications may be made withoutdeviating from the disclosure. For example, the dimensions described inthis disclosure may be modified without changing the general sizingrelationships between various components. Additionally, materials knownto a person of ordinary skill in the art that are not explicitly listedmay be used instead of the materials explicitly described.

Aspects of the disclosure described in the context of particularembodiments may be combined or eliminated in other embodiments. Forexample, features of manufactured seeds described as being useful foragricultural embryos may be combined with features of manufactured seedsdescribed as being useful for tree embryos. Additionally, orientationsof embryos shown with particular embodiments of shoot restraints may beused with other embodiments.

Further, while advantages associated with certain embodiments of thedisclosure may have been described in the context of those embodiments,other embodiments may also exhibit such advantages, and not allembodiments need necessarily exhibit such advantages to fall within thescope of the disclosure. For example, manufactured seeds having onlycoated seal assemblies according to the disclosure may or may not haveall of the advantages of manufactured seeds having parabolic shootrestraints, parabolic seal assemblies, and treated seal assembliesaccording to the disclosure. Accordingly, the invention is not limitedexcept as by the appended claims.

EXAMPLES

The following examples will serve to illustrate aspects of the presentdisclosure. The examples are intended only as a means of illustrationand should not be construed to limit the scope of the disclosure in anyway. Those skilled in the art will recognize many variations that may bemade without departing from the spirit of the disclosure. Certainterminology used in the disclosure are defined as follows:

“Grew between seal assembly” refers to an abnormal growth patterncharacterized by an embryo that elongates to grow between the primaryend seal and the secondary end seal in a manufactured seed.

“Grew through” refers to a germinant that emerges through a secondaryend seal but part of the end seal is still attached to the hypocotoyl.

“Lateral root” means a secondary root or a root that has its originsfrom the primary root. The presence of a lateral root generallyindicates health and vigor of the embryo.

“Normalcy” with respect to embryo germination denotes the presence ofall plant parts (radicle, hypocotyls, cotyledon(s), epicotyls) and noabnormal growth at time of evaluation. Generally the radicle should beat least 3 mm long after at least 24 days germination to denotenormalcy. A “normal germinant” has all of the plant parts describedabove, a radicle with a length greater than 3 mm after at least 24 daysgermination, and no visibly and immediately discernable abnormal growthpatterns.

“Pretzeled hypocotyls” refers to a type of abnormal growth characterizedby abnormally looped, twisted, and/or kinked hypocotyls. This type ofabonormal growth is often associated with mechanical resistance toelongation during germination.

“Root end in air” refers to a type of abnormal growth characterized byan inverted orientation of the root end. This type of abnormal growth isthought occur when the geotropic sensing organ of the root is damaged ormissing.

“Root in cavity” refers to a condition where the root end of thegerminant fails to emerge from the seed or grows back into the cavityupon germination.

Generally, the types of abnormal growth observed in the Examples do notindefinitely indicate that there is a germination problem that will befatal to the embryos. However, noting the quantity and quality ofabnormal growth patterns is a reliable way to comparatively evaluate thevarious methods and means employed for making manufactured seeds.Fortunately, plant embryonic tissue is exquisitely sensitive tonon-natural conditions and manifests that sensitivity in waysdiscernable to a trained observer.

Example 1 Breaking Strength of Conventionally Treated Seal Assemblies

In a first example, seal assemblies were treated with variousconventional coatings and the resulting breaking strength were measured.For this experiment, manufactured seeds having conventional sealassemblies and conventional shoot restraints were provided. Secondaryend seals were provided and treated with six different coatings (appliedwith a paintbrush) and tested 18 hours after application to determinebreaking strength. The six different coatings were: (1) no coating; (2)petroleum jelly; (3) a mixture of petroleum jelly and triple antibioticointment; (4) silicone grease; (5) lanolin heated to 37° C.; and (6)lanolin heated to 50° C. Approximately 30 specimens of each type weretested.

The secondary end seals were made from Parafilm® manufactured byAmerican Can Company, Chicago, Ill. and supplied by VWR (catalog#52858-032). The end seals were each stretched to a diameter of about0.10 inches prior to application of the coatings. The triple antibioticointment purchased over-the-counter from Rite Aid™ Pharmacies. Thelanolin product number L-7387 purchased from Sigma Chemical Company. Thesilicone grease manufactured by Dow Corning, Compound 4 ElectricalInsulating. FIG. 27 summarizes the results.

Example 2 Breaking Strength of Seal Assemblies Treated According toEmbodiments of the Disclosure

In a second example, the breaking strength of seal assemblies treatedwith paraffin oil according to embodiments of the disclosure werecompared with conventional seal assemblies treated with conventionalsubstances. For this experiment, secondary end seals were constructed ina manner similar to that described in Example 1. The secondary end sealswere treated with paraffin oil, treated with Petroleum Jelly (Vaselinebrand), treated with triple-antibiotic ointment, treated with a mixtureof the previous components, or left untreated. Approximately 30specimens of each type were tested. Some of the specimens were stored atroom temperature while the treatment softened the end seal for up toseven days. FIG. 28 summarizes the resulting breaking strengths of eachspecimen.

Example 3 Impact of Paraffin Oil Level on Germination

In a third example, seal assemblies were treated with paraffin oilaccording to embodiments of the disclosure to evaluate the resultingimpact on germination. For this experiment, manufactured seeds havingconventional end seal assemblies and conventional shoot restraints wereprovided. The secondary end seals were treated with varying amounts ofparaffin oil according to the treatment schedule in Table 2.

TABLE 2 Treatment Schedule for Example 3 Treatment Amount of Number ofNumber Paraffin Oil Seeds T1 0.1 mg 70 T2 0.3 mg 72 T3 0.9 mg 67 T4 1.3mg 87

After the paraffin oil was applied to the secondary end seal, themanufactured seeds were stored overnight in a refrigerator. A tertiaryend seal was then attached to the secondary end seal, the seeds weresown in a non-sterile environment, and germination was observed for 43days. FIG. 29 shows the percentage of the population in each treatmentschedule that achieved full germination or partial germination.

This experiment provided evidence to support the hypotheses that toomuch paraffin oil on seal assemblies may inhibit germination. The T3treatment (0.9 mg) exhibited the best performance. At day 43, the T4treatment (1.3 mg) showed nearly equivalent performance to the T1treatment (0.1 mg).

Example 4 Impact of Paraffin Oil Level and Pin Depth on BreakingStrength

In a fourth example, seal assemblies were treated with paraffin oilaccording to embodiments of the disclosure to evaluate the impact ofincreased oil levels on breaking strength. For this experiment,secondary end seals were constructed in a manner similar to thatdescribed in Examples 1 and 2. Prior to application of the paraffin oil,the secondary end seals were pre-stressed using pins to varying depths:0.118 inches, 0.157 inches, and 0.192 inches. The secondary end sealswere then treated with varying amounts of paraffin oil according to thetreatment schedule in Table 3.

TABLE 3 Treatment Schedule for Example 4 Treatment Amount of NumberParaffin Oil Pin Depth T5 1.6 mg .118 inches T6 3.2 mg .118 inches T71.6 mg .157 inches T8 3.2 mg .157 inches T9 1.6 mg .177 inches T10 3.2mg .177 inches T11 1.6 mg .192 inches T12 3.2 mg .192 inches

After the paraffin oil was applied to the secondary end seal, breakingstrength was measured at 24 hours and 48 hours respectively. FIG. 30shows representative results of the measurements at 24 hours. FIG. 31presents the results of the measurements at 48 hours.

Example 5 Impact of Paraffin Oil Level on Breaking Strength,Germination, and Root Damage

In a fifth example, seal assemblies were treated with paraffin oilaccording to embodiments of the disclosure to evaluate the impact ofincreased oil levels on breaking strength, germination, and root damage.For this experiment, manufactured seeds having conventional sealassemblies and conventional shoot restraints were provided. Prior toapplication of the paraffin oil, the secondary end seals werepre-stressed using pins to a depth of 0.192 inches. The secondary endseals were then treated with varying amounts of paraffin oil accordingto the treatment schedule in Table 4.

TABLE 4 Treatment Schedule for Example 5 Treatment Amount of NumberParaffin Oil T13 1.6 mg T14 3.2 mg T15 6.4 mg

Approximately 100 specimens for each treatment type (T13, T14, and T15)were sown in a sterile environment and allowed to germinate for 32 days.During this time, various properties indicative of abnormal growth orhealthy germination were observed and scored. The results are summarizedin Tables 5-8 below.

TABLE 5 Average Root Length (scored at 32 days) Radicle HypocotylsCotyledon Epicotyls Length (mm) Length (mm) Length (mm) Length (mm) T1332.6 25.70 20.54 11.27 T14 39.19 28.97 20.56 10.83 T15 25.86 23.84 20.679.57

Table 5 shows that the average radicle length for the 3.2 mg oiltreatment (T14 treatment) was 39 mm compared to 25 mm for the 6.4 mg oiltreatment (T15 treatment). It seems that 3.2 mg oil treatment (T14treatment) allowed good root growth where the 6.4 mg oil treatment (T15treatment) may have been too much oil resulting in inhibited growth.

TABLE 6 Percent Above/Below Germination (scored at 32 days) 5 2 4Lateral 1 Partial 3 Root End in Root Full Germ Germ No Germ Air PresenceT13 38.68% 38.05% 19.23% 4.02% 52.51% T14 42.39% 42.39% 13.17% 2.02%58.43% T15 28.63% 44.92% 21.34% 5.09% 44.12%

Table 6 shows a grouping of the seeds in Example 5 according to fivenormalcy categories: (1) full germination; (2) partial germination; (3)no germination; (4) root end in air; and (5) lateral root presence. Theresults Table 6 show a trend similar to Table 5. The 3.2 mg oiltreatment (T14 treatment) performed the best with 42% full germinants.Table 5 shows that while the 6.4 mg oil treatment (T15 treatment) had28% full germinants, it also had 44% partial germinants.

TABLE 7 Percent Above/Below Germination (scored at 32 days) 2 Normal if5 1 Fully 3 4 Root in Normal Extracted Not Normal Unchanged Cavity T1361.66% 6.10% 21.90% 7.25% 3.07% T14 67.61% 4.06% 21.33% 4.97% 2.00% T1554.12% 5.12% 27.62% 7.06% 6.06%

Table 7 shows a grouping of the seeds in Example 5 the seeds accordingto five normalcy categories: (1) normal germinants; (2) would be normalif fully extracted from seed; (3) not normal; (4) unchanged (no visibleelongation); and (5) root in cavity. Table 7 shows the same trend withthe 3.2 mg oil treatment (T14 treatment) having the best percent ofnormalcy at 67%. The 6.4 mg oil treatment (T15 treatment) had only 54%normalcy which is still well below the 3.2 mg oil treatment (T14treatment). Each treatment had at least 20% in the “not normal”category.

TABLE 8 Percent with Seal Assembly Problems (scored at 32 days) 1 2 3 4No Grew Through, Grew Between Pretzeled Problems Stuck to Hypo SealAssembly Hypo T13 59.03% 14.83% 2.91% 5.11% T14 57.55% 20.22% 2.00%4.00% T15 48.88% 17.44% 5.11% 5.11%

Table 8 shows a grouping of the seeds in Example 5 according to fourcategories indicating observable problems with the end seal assembly:(1) no problems; (2) grew through lid, lid is stuck to hypocotyls; (3)grew between seal assembly; (and (4) pretzeled hypocotyls. The 1× oiltreatment (T13 treatment) had the least amount of seal assembly problemswith 59% in the “no problems category. The 3.2 mg oil treatment (T14treatment) was close being with 57% without problems. Table 8 also showsthat all treatments had problems with the hypocotyls getting caught onthe seal assemblies on their way out of the seed.

Example 6 Breaking Strength of Conventional Seal Assemblies and SealAssemblies According to Embodiments of the Disclosure

In a sixth example seal assemblies were treated with paraffin oilaccording to embodiments of the disclosure and conventional treatment toevaluate the impact on breaking strength. For this experiment, secondaryend seals were constructed in a manner similar to that described inExamples 1, 2, and 4. Seven different treatment configurations wereapplied to the secondary end seals as summarized in Table 9.

TABLE 9 Configurations for Example 6 Paraffin Oil Paraffin Oil SealAssembly Coating Amount T16 Conventional (dome) Untreated N/A T17Conventional (dome) Treated 1.6 mg T18 Parabolic dimple Untreated N/AT19 Parabolic dimple Treated 1.6 mg T20 Parabolic dimple Treated 3.2 mgT21 Parabolic dimple Treated 6.4 mg T22 Parabolic dimple Treated 8.0 mg

Breaking strengths of the seal assemblies were measured at various timesranging from 24 hours to 2 weeks. The results are shown in FIG. 32.

Example 7 Germination of Manufactured Seeds According to Embodiments ofthe Disclosure

In a seventh example, germination of manufactured seeds having sealassemblies and parabolic cavities according to embodiments of thedisclosure were compared to conventional manufactured seeds. All sealassemblies were coated with paraffin oil according to embodiments of thedisclosure. For this experiment, four different configurations weretested as summarized in Table 10.

TABLE 10 Configurations for Example 7 Paraffin Oil Seal Assembly CavityCoating T23 Parabolic dimple Parabolic 8.0 mg T24 Parabolic dimpleConventional 8.0 mg T25 Conventional Conventional 1.6 mg

The seeds were sown in a sterile environment and allowed to germinate.Results are shown in FIGS. 33-35. FIG. 33 shows effectiveness ofcotyledon extraction, FIG. 34 shows extraction hang-ups, and FIG. 35shows normalcy. In this experiment, seal assemblies according toembodiments of the disclosure and conventional cavities significantlyreduced lid hang-ups down to 6% and eliminated grow-betweens which havebecome a problem with other germination studies.

We claim:
 1. A manufactured seed comprising: a seed shell comprising astructure having an open end and a closed end; a restraint disposedwithin the seed shell, the restraint comprising a parabolic cavity; anda seal assembly disposed on the open end of the seed shell in aconfiguration effective to seal the seed shell.
 2. The manufactured seedof claim 1, further comprising an embryo disposed within the paraboliccavity.
 3. The manufactured seed of claim 2 wherein the embryo isdisposed in the parabolic cavity in a substantially perpendicularconfiguration with respect to a plane in which the sealing assembly isarranged.
 4. The manufactured seed of claim 3 wherein the embryocomprises a root end and a shoot end and the embryo is disposed in theparabolic cavity shoot end first.
 5. The manufactured seed of claim 2wherein the embryo is disposed in the parabolic cavity in asubstantially parallel configuration with respect to a plane in whichthe sealing assembly is arranged.
 6. The manufactured seed of claim 5wherein the secondary end seal comprises one or more parabolic dimples.7. The manufactured seed of claim 2 wherein the embryo is disposed inthe parabolic cavity in a substantially skew configuration with respectto a plane in which the sealing assembly is arranged.
 8. Themanufactured seed of claim 7 wherein the one or more parabolic dimplesare coated, at least partially, in a paraffin oil.
 9. The manufacturedseed of claim 2 wherein the embryo comprises a tree embryo.
 10. Themanufactured seed of claim 2 wherein the embryo comprises anagricultural crop embryo.
 11. The manufactured seed of claim 1 whereinthe open end of the seed shell has a seed shell diameter D1 and therestraint further comprises: an upper portion having an upper diameterD2, the seed shell diameter D1 being larger than the upper diameter D2;and a lower portion having a lower diameter D4, the upper diameter D2being larger than the lower diameter D4.
 12. The manufactured seed ofclaim 11 wherein the parabolic cavity has a cavity diameter D5, thecavity diameter D5 being at least as large as the lower diameter D4. 13.The manufactured seed of claim 12 wherein the upper restraint portionhas an upper depth L2, and the parabolic cavity has a cavity depth L5,the cavity depth L2 being substantially larger than the upper depth L5.14. The manufactured seed of claim 13 wherein the cavity depth L5 isapproximately 0.10 inches to approximately 0.20 inches.
 15. Themanufactured seed of claim 13 wherein the upper depth L2 isapproximately 0.15 to approximately 0.30 inches.
 16. The manufacturedseed of claim 13 wherein the upper depth L2 is approximately 0.20 toapproximately 0.30 inches.
 17. The manufactured seed of claim 11 whereinthe seed shell has a seed shell diameter D1 and the parabolic cavity hasa cavity diameter D5, the cavity diameter D5 being substantially thesame size as the seed shell diameter D1.
 18. The manufactured seed ofclaim 17 wherein the seed shell diameter D1 is approximately 0.30 inchesto approximately 0.40 inches, the upper diameter D2 is approximately0.20 inches to approximately 0.35 inches, the lower diameter D4 isapproximately 0.10 to 0.15 inches, and the cavity diameter D5 isapproximately 0.10 inches to approximately 0.30 inches.
 19. Themanufactured seed of claim 17 wherein the seed shell diameter D1 isapproximately 0.20 inches to approximately 0.30 inches, the upperdiameter D2 is approximately 0.15 to approximately 0.25 inches, thelower diameter D4 is approximately 0.10 to approximately 0.18 inches andthe cavity diameter D5 is approximately 0.10 to approximately 0.20inches
 20. The manufactured seed of claim 1 wherein the seal assemblyfurther comprises a primary end seal and a secondary end seal.